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Lipids & Membranes
Copyright © 1999-2006 by Joyce J. Diwan. All rights reserved.
Lipids are non-polar (hydrophobic) compounds, soluble in organic solvents. Most membrane lipids are amphipathic, having a non-polar end and a polar end. Fatty acids consist of a hydrocarbon chain with a carboxylic acid at one end.
A 16-C fatty acid: CH3(CH2)14-COONon-polar polar
A 16-C fatty acid with one cis double bond between C atoms 9-10 may be represented as 16:1 cis D9.
Double bonds in fatty a 3 acids usually have the 1 O 4 2 cis configuration. Most naturally fatty acid with a cis-D9 occurring fatty acids double bond have an even number of carbon atoms. Some fatty acids and their common names: 14:0 myristic acid; 16:0 palmitic acid; 18:0 stearic acid; 18:1 cisD9 oleic acid 18:2 cisD9,12 linoleic acid 18:3 cisD9,12,15 a-linonenic acid 20:4 cisD5,8,11,14 arachidonic acid 20:5 cisD5,8,11,14,17 eicosapentaenoic acid (an omega-3)
fatty acid with a cis-D9 double bond
There is free rotation about C-C bonds in the fatty acid hydrocarbon, except where there is a double bond. Each cis double bond causes a kink in the chain. Rotation about other C-C bonds would permit a more linear structure than shown, but there would be a kink.
Formation of an ester: O R'OH + HO-C-R" CH2OH glycerol O R'-O-C-R'' + H2O . CH2OH H C OH Hydroxyls at C1 & C2 are esterified to fatty acids. They have a glycerol backbone. with loss of H2O.Glycerophospholipids Glycerophospholipids (phosphoglycerides). An ester forms when a hydroxyl reacts with a carboxylic acid. are common constituents of cellular membranes.
.Phosphatidate O O R1 C O H2C CH H2C O O C O P O O R2 phosphatidate In phosphatidate: fatty acids are esterified to hydroxyls on C1 & C2 the C3 hydroxyl is esterified to Pi.
The 2 fatty acids tend to be non-identical.O O R1 C O H2C CH H2C O O C O P O O X R2 glycerophospholipid In most glycerophospholipids (phosphoglycerides). . or inositol. They may differ in length and/or the presence/absence of double bonds. Pi is in turn esterified to OH of a polar head group (X): e. choline. ethanolamine. serine.g. glycerol..
with inositol as polar head group.O O R1 C O H2 C CH H2 C O O C O P O OH H OH OH H H H OH O H OH R2 phosphatidylinositol H Phosphatidylinositol. In addition to being a membrane lipid. is one glycerophospholipid. phosphatidylinositol has roles in cell signaling. .
is another glycerophospholipid. with choline as polar head group.O O R1 C O H2C CH H2C O O C O P O O CH2 CH2 R2 CH3 + N CH3 CH3 phosphatidylcholine Phosphatidylcholine. It is a common membrane lipid. .
& the polar head group (X) H2C O CH H2C O C O R2 R1 C O P O O X glycerophospholipid polar "kink" due to double bond non-polar hydrocarbon tails of fatty acids (R1. Pi. R2). non-polar .O O Each glycerophospholipid includes a polar region: glycerol. carbonyl O of fatty acids.
sphingosine-1-P CH3 . Other derivatives of sphingosine are commonly found as constituents of biological membranes. and a polar domain that includes an amino group.Sphingolipids are derivatives of the lipid sphingosine. O OH H2C H C H3N+ OH CH CH HC (CH2 )12 O P O H2C O sphingosine OH H C H3N+ CH CH HC (CH2 )12 CH3 Sphingosine may be reversibly phosphorylated to produce the signal molecule sphingosine-1-phosphate. which has a long hydrocarbon tail.
to yield a ceramide. . OH H2C H C NH O C R OH CH CH HC (CH2 )12 CH3 sphingosine CH3 ceramide In the more complex sphingolipids. a polar “head group" is esterified to the terminal hydroxyl of the sphingosine moiety of the ceramide.OH H2C H C H3N+ OH CH CH HC (CH2 )12 The amino group of sphingosine can form an amide bond with a fatty acid carboxyl.
is similar in size and shape to the glycerophospholipid phosphatidyl choline. CH3 phosphocholine sphingosine O Sphingomyelins are common constituent of plasma membranes fatty acid Sphingomyelin Sphingomyelin. . with a phosphocholine head group.CH3 H3C N + O H2 C H2 C O P O H2C H C NH C R O OH CH CH HC (CH2 )12 CH3 Sphingomyelin has a phosphocholine or phosphethanolamine head group.
are commonly found in the outer leaflet of the plasma membrane bilayer. with their sugar chains extending out from the cell surface. collectively called glycosphingolipids. including the acidic sugar derivative sialic acid. R (CH2 )12 cerebroside with A ganglioside is a -galactose head group CH3 ceramide with a polar head group that is a complex oligosaccharide. .CH2OH A cerebroside is a O OH H sphingolipid OH O H OH H (ceramide) with a H H2C C CH H monosaccharide H OH NH CH such as glucose or galactose as polar O C HC head group. Cerebrosides and gangliosides.
. a spherical micelle is a stable configuration for amphipathic lipids with a conical shape. possible molecular arrangements: Various micelle structures. . E.g. Bilayer Spherical Micelle Depending on the lipid. such as fatty acids. A bilayer. such as phospholipids. This is the most stable configuration for amphipathic lipids with a cylindrical shape.Amphipathic lipids in association with water form complexes in which polar regions are in contact with water and hydrophobic regions away from water.
packing is highly ordered.Membrane fluidity: The interior of a lipid bilayer is normally highly fluid. due to cis double bonds. Kinks in fatty acid chains. and lower the phase transition temperature. hydrocarbon chains of phospholipids are disordered and in constant motion. At lower temperature. a membrane containing a single phospholipid type undergoes transition to a crystalline state in which fatty acid tails are fully extended. . interfere with packing in the crystalline state. liquid crystal crystal In the liquid crystal state. & van der Waals interactions between adjacent chains are maximal.
PDB 1N83 cholesterol . HO Cholesterol But it has one polar group. making it amphipathic. a hydroxyl. Cholesterol is largely hydrophobic.Cholesterol. an important constituent of cell membranes. has a rigid ring system and a short branched hydrocarbon tail.
The OH group of cholesterol forms hydrogen bonds with polar phospholipid head groups. .HO Cholesterol Cholesterol in membrane Cholesterol inserts into bilayer membranes with its hydroxyl group oriented toward the aqueous phase & its hydrophobic ring system adjacent to fatty acid chains of phospholipids.
.Interaction with the relatively rigid cholesterol decreases the mobility of hydrocarbon tails of phospholipids. Cholesterol in membrane But the presence of cholesterol in a phospholipid membrane interferes with close packing of fatty acid tails in the crystalline state. and thus inhibits transition to the crystal state. Phospholipid membranes with a high concentration of cholesterol have a fluidity intermediate between the liquid crystal and crystal states.
In the absence of cholesterol. that include many lipids with long-chain saturated fatty acids. The inner mitochondrial membrane lacks cholesterol. such membranes would crystallize at physiological temperatures.Two strategies by which phase changes of membrane lipids are avoided: Cholesterol is abundant in membranes. but includes many phospholipids whose fatty acids have one or more double bonds. . which lower the melting point to below physiological temperature. such as plasma membranes.
The apparent constraints on lateral movements of lipids (and proteins) has been attributed to integral membrane proteins. . functioning as a picket fence.Lateral mobility of a lipid. anchored to the cytoskeleton. See the website of the Kusumi laboratory. Lateral Mobility High speed tracking of individual lipid molecules has shown that lateral movements are constrained within small membrane domains. is depicted at right and in an animation. within the plane of a membrane. Hopping from one domain to another occurs less frequently than rapid movements within a domain.
The two leaflets of a bilayer membrane tend to differ in their lipid composition. Flip Flop Flip-flop would require the polar head-group of a lipid to traverse the hydrophobic core of the membrane. .Flip-flop of lipids (from one half of a bilayer to the other) is normally very slow. Some membranes contain enzymes that actively transport particular lipids from one monolayer to the other. Flippases catalyze flip-flop in membranes where lipid synthesis occurs.
. e.g. and/or chelators that bind divalent cations. with mostly hydrophilic surfaces. change of pH.. Often peripheral proteins can be dislodged by conditions that disrupt ionic & H-bond interactions.peripheral Membrane proteins may be classified as: peripheral integral having a lipid anchor lipid anchor lipid bilayer integral Membrane Proteins Peripheral proteins are on the membrane surface. extraction with solutions containing high concentrations of salts. They are water-soluble.
. The enzymes that create or degrade these lipids are subject to signal-mediated regulation.hypothetical protein regulatory catalytic membrane domain domain binding Many proteins have a modular design. providing a mechanism for modulating affinity of a protein for a membrane surface. with different segments of the primary structure folding into domains with different functions. Some cytosolic proteins have domains that bind to polar head groups of lipids that transiently exist in a membrane.
O O R1 C O H2C CH H2C O O C O P O OH 2 1 6 R2 O H OPO32 5 H OH 3 OH H 4 PIP2 H phosphatidylinositol4.g. .5-P2). pleckstrin homology (PH) domains bind to phosphorylated derivatives of phosphatidylinositol. Some PH domains bind PIP2 (PI-4.5-bisphosphate H H OPO32 E..
. PI-3-P.O O R1 C O H2C CH H2C O O C O P O O H 1 6 R2 phosphatidylinositol3-phosphate OH 2 OH 5 OH H OPO32 H 3 4 H H H OH Other pleckstrin homology domains recognize and bind phosphatidylinositol derivatives with Pi esterified at the 3' OH of inositol.g.4. PI-3.5-P3. PI-3. E.4-P2..
g. A protein may be released from plasma membrane to cytosol via depalmitoylation. A protein may link to the cytosolic surface of the plasma membrane via a covalently attached fatty acid (e. . palmitate or myristate) or an isoprenoid group.lipid anchor H3C (CH2)14 O C S O CH2 C CH NH membrane cysteine residue palmitate Some proteins bind to membranes via a covalently attached lipid anchor. that inserts into the bilayer. hydrolysis of the ester link.. Palmitate is usually attached via an ester linkage to the thiol of a cysteine residue.
is attached to some proteins via a thioether linkage to a cysteine thiol. lipid anchor membrane .CH3 H3C C CH CH2 CH2 CH3 C CH CH2 CH3 CH2 C CH CH2 S Protein farnesyl residue linked to protein via cysteine S An isoprenoid such as a farnesyl residue.
The linkage is similar to the following. GPI-linked proteins may be released from the outer cell surface by phospholipases. .Glycosylphosphatidylinositols (GPI) are complex glycolipids that attach some proteins to the outer surface of the plasma membrane. although the oligosaccharide composition may vary: protein (C-term.mannose mannose .phosphoethanolamine – mannose .N-acetylglucosamine – inositol (of PI in membrane) The protein is tethered some distance out from the membrane surface by the long oligosaccharide chain.) .
Intramembrane domains have largely hydrophobic surfaces. Often they span the bilayer. that interact with membrane lipids. .peripheral lipid anchor lipid bilayer integral Membrane Proteins Integral proteins have domains that extend into the hydrocarbon core of the membrane.
purified integral proteins tend to aggregate & come out of solution. coating hydrophobic surfaces of integral proteins. Polar domains of detergents interact with water. . If detergents are removed. Their hydrophobic surfaces associate to minimize contact with water. detergent solubilization polar non-polar Protein with bound detergent Hydrophobic domains of detergents substitute for lipids.membrane Amphipathic detergents are required for solubilization of integral proteins from membranes.
Membrane fragments assumed to be lipid rafts are found to be resistant to detergent solubilization. which has facilitated their isolation & characterization. .Lipid rafts: Complex sphingolipids tend to separate out from glycerophospholipids & co-localize with cholesterol in membrane microdomains called lipid rafts.
Santini & coworkers). • See diagram (in article by J. . Lipid raft domains tend to be thicker than adjacent membrane areas. • Glycerophospholipids often include at least one fatty acid that is kinked. in part because the saturated hydrocarbon chains of sphingolipids are more extended. due to one or more double bonds. Differences in molecular shape may contribute to a tendency for sphingolipids to separate out from glycerophospholipids in membrane microdomains. • Sphingolipids usually lack double bonds in their fatty acid chains.
CH3 H3C N + O H2 C H2 C O P O H2C H C NH C R O OH CH CH CH3 phosphocholine sphingosine O HO HC (CH2 )12 CH3 Cholesterol fatty acid Sphingomyelin Hydrogen bonding between the hydroxyl group of cholesterol and the amide group of sphingomyelin may in part account for the observed affinity of cholesterol for sphingomyelin in raft domains. .
• Otherwise soluble signal proteins often assemble in complexes at the cytosolic surface of the plasma membrane in part via insertion of attached fatty acyl or isoprenoid lipid anchors into raft domains. . • Integral proteins may concentrate in raft domains via interactions with raft lipids or with other raft proteins. • Some raft domains contain derivatives of phosphatidylinositol that bind signal proteins with pleckstrin homology domains. Proteins involved in cell signaling often associate with lipid raft domains.
caveolae cytosol Caveolin is a protein associated with the cytosolic leaflet of the plasma membrane in caveolae. Electron micrograph & information about caveolae (home page of Deborah Brown at SUNY Stony Brook). Diagram & information about lipid rafts (website of Maciver lab at University of Edinburgh). . Caveolin interacts with cholesterol and self-associates as oligomers that may contribute to deforming the membrane to create the unique morphology of caveolae. Caveolae are invaginated lipid raft domains of the plasma membrane that have roles in cell signaling and membrane internalization.
Integral proteins are difficult to crystallize for X-ray analysis. detergents must be present during crystallization. C membrane N . A membrane-spanning a-helix is the most common structural motif found in integral proteins. Because of their hydrophobic transmembrane domains.Integral protein structure Atomic-resolution structures have been determined for a small (but growing) number of integral membrane proteins.
Colors: C N O R-group (H atoms not shown).a-helix R-groups in magenta In an a-helix. The largely hydrophobic R-groups of a membranespanning a-helix contact the hydrophobic membrane core. . while the more polar peptide backbone is buried. amino acid R-groups protrude out from the helically coiled polypeptide backbone.
isoleucine. valine) predominate in the middle of the bilayer. alanine. I) H C COO leucine (Leu. V) H H3N+ C COO COO H3N+ CH CH3 CH2 CH3 CH CH3 CH3 amino acids: non-polar aliphatic R-groups Particular amino acids tend to occur at different positions relative to the surface or interior of the bilayer in transmembrane segments of integral proteins. L) H H3N+ C CH2 CH CH3 CH3 COO valine (Val. .alanine (Ala. Residues with aliphatic side-chains (leucine. A) H H3N+ C CH3 isoleucine (Ile.
suit them for localization at the polar/apolar interface. HN OH It has been suggested that the polar character of the tryptophan amide group and the tyrosine hydroxyl. . along with their hydrophobic ring structures.tryptophan H H2N C CH2 COO tyrosine H H3N+ C CH2 COO Tyrosine and tryptophan are common near the membrane surface.
. with the positively charged groups at the ends of their aliphatic side chains extending toward the polar membrane surface.lysine H H3N+ C CH2 CH2 CH2 CH2 arginine H COO H3N+ C CH2 CH2 CH2 NH COO NH3 C NH2 NH2 Lysine & arginine are often at the lipid/water interface.
Explore with Chime the a-helix colored green at far left.membrane Cytochrome oxidase dimer (PDB file 1OCC) Cytochrome oxidase is an integral protein whose intramembrane domains are mainly transmembrane a-helices. .
Hydropathy plots alone are not conclusive. . Putative hydrophobic transmembrane a-helices have been identified this way in many membrane proteins.A 20-amino acid a-helix just spans a lipid bilayer. Hydropathy plots are used to search for 20-amino acid stretches of hydrophobic amino acids in the primary sequence of a protein for which a crystal structure is not available. Protein topology studies are used to test the transmembrane distribution of protein domains predicted by hydropathy plots.
If two transmembrane a-helices are predicted.C membrane N C N If a hydropathy plot indicates one 20-amino acid hydrophobic stretch (1 putative transmembrane a-helix). N & C termini should be on the same side. . The segment between the a-helices should be on the other side. topology studies are expected to confirm location of N & C termini on opposite sides of membrane.
Degradation a protein segment indicates exposure to the aqueous phase on the side of the membrane to which a protease is added. For example: Protease enzymes. . Binding indicates surface exposure of a protein segment on the side to which the Ab is added.Transmembrane topology is tested with impermeant probes. Such studies have shown that all copies of a given type of integral protein have the same orientation relative to the membrane. Flip-flop of integral proteins does not occur. Monoclonal antibodies raised to peptides equivalent to individual segments of the protein. added on one side of a membrane.
A “helical wheel” looks down the axis of an a-helix. . Polar amino acid R-group Non-polar amino acid R-group An a-helix lining a water-filled channel might have polar amino acid R-groups facing the lumen. & non-polar R-groups facing lipids or other hydrophobic a-helices. Such mixed polarity would prevent detection by a hydropathy plot. projecting sidechains onto a plane.Simplified helical wheel diagram of four a-helices lining the lumen of an ion channel.
PDB 1AOS . A barrel is a sheet rolled up to form a cylindrical pore. a family of bacterial outer envelope channel proteins called porins have instead barrel structures. Porin Monomer At right is shown one channel of a trimeric porin complex.Porin -barrel While transmembrane ahelices are the most common structural motif for integral proteins.
. polar R group. Much of porin primary structure consists of alternating polar & non-polar amino acids. amino acid R-groups alternately point above & below the sheet. • Polar residues face the aqueous lumen. non-polar R group In a -sheet. • Non-polar residues are in contact with membrane lipids. Explore an example of a bacterial porin with Chime.